Fast and simple reaction setup

A single-tube assay setup allows for routine automation, and delivers genotypes with only 90 minutes of cycling time (Figure 1). Hot-start enzymes enable benchtop reaction setup, and stability of reactions for up to 3 days before and after cycling at room temperature.

Figure 1. Simple, one-tube reaction chemistry supports streamlined lab processes. All reagents are combined in the initial reaction setup that is stable for up to 48 hours at room temperature. Reaction setup, instrument run-time, and data acquisition may be completed in under 3 hours.

Superior chemistry for SNP detection

Superior discrimination versus traditional methods. Blocked primers minimize non-specific amplification. The 3' end of rhAmp primers incorporate a blocking group that prevents extension unless cleavage and de-blocking occur by RNase H2 enzyme. RNase H2 enzyme recognizes this RNA base only if it is hybridized to its perfect complement, initiating primer cleavage and activation.

Figure 2. Schematic representation of a rhAmp SNP Genotyping PCR cycle. All components needed to measure both alleles are combined in a single reaction before cycling. 1) Both allele-specific primers query the SNP locus. 2) RNase H2 enzyme cleaves the primers that are perfectly annealed to the target sequence, removing the RNA base and 3′ blocking modification, which allows extension by the IDT Taq Polymerase. 3) During the first two amplification cycles, a tail sequence is incorporated into the amplicon that is subsequently recognized by a universal, probe-based reporter system. 4) Polymerase extension leads to degradation of the probe and signal generation.